I International Journal of Innovations in Agricultural Sciences (IJIAS) Journal of In

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1 Available online at Volume 1; Issue - 1; Year 2017; Page: ISSN: DOI: /ijias I International Journal of Innovations in Agricultural Sciences (IJIAS) Journal of In PRODUCTION OF BIODIESEL BY MICROALGAE AND MACROALGAE FROM MANGROVES OF PICHAVARAM K. Sivakumar*, G. Kumaresan and K. Kesavamoorthy, Department of Agricultural Microbiology, Faculty of Agriculture, Annamalai University, Annamalai Nagar , Tamilnadu, India. Abstract Biodiesel, as an alternative fuel, has many benefits. It is biodegradable, non-toxic and compared to petroleum-based diesel, has a more favorable combustion emission profile, such as low emissions of carbon monoxide, particulate matter and unburned hydrocarbons. In brief, these merits make biodiesel a good alternative to petroleum based fuel. Biodiesel feedstock derived from microalgae and macroalgae have emerged as one of the most promising alternative sources of lipid for use in biodiesel production because of their high photosynthetic efficiency to produce biomass and their higher growth rates and productivity compared to conventional crops. In addition to their fast reproduction, they are easier to cultivate than many other types of plants and can produce a higher yield of oil for biodiesel production. In this present, work biodiesel was produced using the species of microalgae Chlorella emersonii and Botrycoccus braunii due to its high oil content. Biodiesel productions through macroalgae oil are in preliminary phase. Therefore, results and methodology will not be presented in this work. Technological assessment of process was carried out to evaluate their technical benefits, limitations and quality of final product. In this work, biodiesel from microalgae oil was produced by an alkali-catalyzed transesterification and it was achieved 93 % of mass conversion. Results show that all parameters analyzed meet the standard and legislation requirements. This evidence proves that in those operational conditions the biodiesel produced from microalgae can substitute petroleum-based diesel. Key words: Biodiesel, Algae, Pichavaram, Chlorella emersonii and Botrycoccus braunii. 1. Introduction Energy is the most fundamental requirement for human existence and activities. As an effective fuel, petroleum has been serving the world to meet its need of energy consumption. But the dependence of mankind entirely on the fossil fuels could cause a major deficit in future. The application of biodiesel to our diesel engines for daily activities was advantageous for its environmental friendliness over petro-diesel (Atadashi et al., 2010). The main advantages of using biodiesel is that it is biodegradable, can be used without modifying existing engines, and produces less harmful gas emissions such as sulfur oxide (Chand 2002; Atadashi et al., 2010). Biodiesel reduces net carbon-dioxide emissions by 78 % on a life cycle basis when compared to conventional diesel fuel (Gunvachai et al., 2007). Puppan (2002) have discussed the advantages of biofuels over fossil fuels to be: (a) availability of renewable sources; (b) representing CO 2 cycle in combustion; (c) environmentally friendly and (d) biodegradable and sustainable. Other advantages of biodiesel are as follows: portability, ready availability, lower sulfur & aromatic content and high combustion characteristics. Biodiesel, which was considered as a possible substitute of conventional diesel fuel is composed of fatty acid methyl esters that can be prepared from triglycerides in vegetable oils by 2017 Published by JPS Scientific Publications Ltd. All rights reserved

2 K. Sivakumar/International Journal of Innovations in Agricultural Sciences (IJIAS), 1(1): transesterification with methanol (Gerpen, 2005). The resulting biodiesel is quite similar to conventional diesel fuel in its main characteristics (Meher et al., 2006). Transesterification is the process by which the glycerides present in fats or oils react with an alcohol in the presence of a catalyst to form esters and glycerol (Banerjee and Chakraborty, 2009; Enweremadu and Mbarawa, 2009; Zabeti et al., 2009). Catalyst increases the rate of the reaction and also the yield. This reaction proceeds well in the presence of some homogeneous catalysts such as potassium hydroxide (KOH)/sodium hydroxide (NaOH) and sulfuric acid, or heterogeneous catalysts such as metal oxides or carbonates (Basha et al., 2009). Depending on the undesirable compounds (especially FFA and water), each catalyst has its advantages and disadvantages. Sodium hydroxide is very well accepted and widely used because of its low cost and high product yield (Amin, 2009). The most common alcohols widely used are methyl alcohol and ethyl alcohol. Among these two, methanol found frequent application in the commercial uses because of its low cost (Enweremadu and Mbarawa, 2009). Vegetable oils are promising feedstocks for biodiesel production since they are renewable in nature, and can be produced on a large scale and environmentally friendly (Basha et al., 2009). However, it may cause some problems such as the competition with the edible oil market, which increases both the cost of edible oils and biodiesel (Janaun and Ellis, 2010; FAO, 2008; Kansedo et al., 2009). For instance, the mass plantation of monoculture plants could benefit the economy of rural population while negatively affecting the water resources and the biodiversity (FAO, 2008). In order to overcome these disadvantages, many researchers are interested in others feedstock as Algae oil (Chand, 2002; Amin, 2009; Brennan et al., 2009; Mata et al., 2010). The aim of this study was to evaluate biodiesel production through microalgae oil, testing different operating conditions and equipment designs for each stage of processing. Technological assessment of this process was carried out to evaluate their technical benefits, limitations and quality of final product. Microalgae have high potentials in biodiesel production compared to other oil crops. First, the cultivation of microalgae does not need much land as compared to others plants (Chisti, 2007). Biodiesel produced from microalgae will not compromise the production of food and other products derived from crops. Second, microalgae grow extremely rapidly and many algal species are rich in oils. For instance, heterotrophic growth of Chlorella can accumulate lipids as high as 55 % of the cell dry weight after 144 hrs of cultivation (Xu et al., 2006). Oil levels of % are common in microalgae (Chisti, 2007). These technological advances suggest that the industrial production of biodiesel from microalgal oils may be feasible in the near future. The advantages of culturing microalgae as a resource of biomass are: a) Algae are considered to be a very efficient biological system for harvesting solar energy for the production of organic compounds. b) Algae are non-vascular plants, lacking (usually) complex reproductive organs. c) Many species of algae can be induced to produce particularly high concentrations of chosen, commercially valuable compounds, such as proteins, carbohydrates, lipids and pigments. d) Algae are microorganisms that undergo a simple cell division cycle. e) Microalgae can be grown using sea or brackish water. f) Algal biomass production systems can easily be adapted to various levels of operational or technological skills (Basha et al., 2009; Amin, 2009; Brennan et al., 2009; Mata et al., 2010; Hossain et al., 2008). The use of alkali catalysts in transesterification reaction of microalgae oil is somewhat limited, because FFA content presents in this feedstock oil reacts with the most common alkaline catalysts (NaOH, KOH and CH 3 ONa) and forms soap (Banerjee and Chakraborty, 2009). This reaction is undesirable because soap lowers the yield of the biodiesel and inhibits the separation of esters from glycerol. In addition, it

3 K. Sivakumar/International Journal of Innovations in Agricultural Sciences (IJIAS), 1(1): binds with the catalyst meaning that more catalyst will be needed and hence the process will involve a higher cost (Enweremadu and Mbarawa, 2009). Feedstocks with high free fatty acid will react undesirably with the alkali catalyst thereby forming soap. Maximum amount of free fatty acids acceptable in an alkali - catalyzed system is below 3 wt. % FFA. If the oil feedstock has a FFA content over 3 wt. %, a pretreatment step is necessary before the transesterification process (FAO, 2008). 2. Materials and Methods Microalgae species were chosen taking into account its content in oil. Therefore, the species of Chlorella emersonii and Botryococcus braunii was selected which were got from Department of Agricultural Microbiology, Annamalai University. Methanol (purity 99.7 %) and sodium hydroxide with purity of 99 %, which were employed as the alkali catalysts in the reaction. Sulfuric acid, which were employed as the acid catalysts in the transesterification reaction. Citric acid, which were employed in the washing process. Microalgae oil The microalgae cultures were development in semi-open flasks, to simulate the conditions of Photobioreactors. The medium used, designated M7 medium was prepared according to seaweed collection. The recovery process for collecting algae biomass was realized trough flocculation by aluminum chloride (AlCl 3.6H 2 O), iron chloride (FeCl.4H 2 O) and aluminum sulfate (Al 2 (SO 4 ) 3.18 H 2 O). For the oil extraction were tested an ultrasound rupture system. Before transesterification, microalgae oil was filtered and heated among C for 30 min. Methanol (5:1 molar ratio methanol/oil) was mixed with sodium hydroxide (0.25 % w/w), until all of the NaOH was dissolved in methanol. This mixture was then added to the oil, and further heated to 60 C for 2 hrs. The ester was purified by washing with distilled water and citric acid and drying at 100 C for 4 hrs. The final polishing process was realized by filtering the methyl ester in a filtering unit system. 3. Results and Discussion Oil extraction For collecting algae biomass it was tested three flocculants: aluminum chloride (AlCl 3.6H 2 O), iron chloride (FeCl.4H 2 O) and aluminum sulfate (Al 2 (SO 4 ) 3.18H 2 O). The results showed a similar behavior for the three flocculants tested. Oil extraction was accomplished by utilization of ultrasound to promote cellular rupture during 1 hour. Table - 1 shows the oil extraction yield for both microalgae species tested. Table 1 Oil extraction yield for Chlorella emersonii and Botryococcus braunii Algae Biomass (g/l) Dry weight (g/l) % Extracted Oil (g/l) (dry weight) % Chlorella emersonii Botryococcus braunii Results showed that Chlorella emersonii achieved 14.8 g oil/l algae culture and Botryococcus braunii obtained 18.3 g oil/l algae culture. Considering the culture time and total algae biomass produced it was reached an extraction coefficient of 3.7 g oil d -1 for Chlorella emersonii and 4.6 g oil d -1 for the Botryococcus braunii. Campbell et al. (2011) realized a technical - economic study for biodiesel production trough Chlorella microalgae. In that study, they concluded that it was necessary an extraction yield of 30 g oil m -2 d -1 to economic ensure the process. The lower growth rate and extraction coefficient obtained in this work can be explained by the utilization of semi-open flasks to simulate an open batch Photobioreactor. According to (Brennan and Owende, 2009) this

4 K. Sivakumar/International Journal of Innovations in Agricultural Sciences (IJIAS), 1(1): type of reactor presents the lowest biomass growth rate. Mass conversion One of the most important dependent variables in this experiment is the mass conversion, which is given by the mass ratio of biodiesel (product) to the total initial mass of the raw material and the additives (Atadashi et al., 2008). Table 2 shows the mass conversion from transesterification reaction on microalgae oil. Mass conversion depends on several parameters like, reaction temperature and pressure, reaction time, rate of agitation, type of alcohol used and molar ratio of alcohol to oil, type and concentration of catalyst used and principle concentration of free fatty acids (FFA) in the feed oil (Banerjee and Chakraborty 2009; Enweremadu and Mbarawa, 2009; Leung et al., 2010). Table 2: Mass conversion (%) of biodiesel from microalgae oil Source FFA content [%] Mass conversion [%] Microalgae oil <1 94 In this case previous FFA determination revealed that microalgae oil has less than 1 %. Therefore, it was realized an alkali - catalyzed transesterification. The results demonstrated that microalgae biodiesel was the maximum mass conversion of 93 %. Quality of Biodiesel The major focal point for biodiesel high quality is the adherence to biodiesel standard specifications. These standard specifications in European Union are established by EN for biodiesel fuel (Atadashi et al., 2008; Banerjee and Chakraborty, 2009; Meher et al., 2006). The results of the analyses in the different types of biodiesel produced (Table - 3) showed that in generally the quality parameters of standard EN was accomplished. Table 3 Results of biodiesel produced from algae oil according to standard EN Microalgae Limits Property Unit biodiesel result Minimum Maximum Test Method Ester content % (m/m) 97,5 ± 0,2 96,5 EN Density at 15 C kg/m ± EN ISO 3675 Viscosity at 40 C mm 2 /s 4,21 ± 0,05 3,00 5,00 EN ISO 3104 Flash point ºC 160 ± 1, EN ISO 3679 Carbon residue % (m/m) 0,05 ± 0,01-0,30 EN ISO Water content mg/kg 472 ± EN ISO 3679 Acid value mg KOH/g 0,36 ± 0,05-0,50 EN Iodine value g iodine100 g 110 ± 1, EN Linolenic acid methyl ester % (m/m) 4,6 ± 0,20-12,0 EN Methanol content % (m/m) 0,1 ±0,01-0,20 EN Group I metals (Na + K) mg/kg 4,02 ± 0,15-5 EN14108

5 K. Sivakumar/International Journal of Innovations in Agricultural Sciences (IJIAS), 1(1): Conclusions Biodiesel is gradually gaining acceptance in the market as an environmentally friendly alternative diesel fuel. However, for biodiesel to established and continue to mature in the market, various aspects must be examined and overcome. Some of the key issues such as improving efficiency of the production process and using low cost feedstock have been reviewed and analyzed in this work. Biodiesel production from microalgae oil presents various advantages compared to other low cost feedstock. However, in other to compete with petroleum-based fuels cost, biodiesel production from microalgae needs to be cheaper. On this work it was possible to conclude the technically feasibility of the process. Microalgae oil had a high yield of conversion and analyses on final biodiesel showed that all quality parameters of the standard have been respected. According to these results, it is possible to conclude that the biodiesel produced with these feedstock was liable to be used in diesel car engines, in a unique way (B100) or blending with fuel diesel (B20, B30 and B50), without decrease of engine efficiency. Nonetheless is important to note that in this work microalgae growth rate and extraction coefficient had low yields which increased cost - production. The solution for this problem requires improvements in algal biology, and in photobioreactor engineering, as the usage of tubular photobioreactors. 5. References 1) Amin, A Review on biofuels oil and gas production process from microalgae. Energy Conversion and Management, 50: ) Atadashi. I. M., M. K. Aroua and A. A. Aziz High quality biodiesel and its diesel engine application: A review. Renewable and Sustainable Energy Reviews, 14: ) Balat, M Current alternative engine fuels. Energy Sources, 27: ) Banerjee, A and R. Chakraborty Parametric sensitivity in transesterification of waste cooking oil for biodiesel production - A review. Resources, Conservation and Recycling, 53: ) Basha, S. A., Gopal, K. R and Jebaraj, S A review on biodiesel production, combustion, emissions and performance. Renewable and Sustainable Energy Reviews, 13: ) Brennan, L and P. Owende Biofuels from microalgae - A review of technologies for production, processing, and extractions of biofuels and co-products. Renewable and Sustainable Energy Reviews (2009). 7) Campbell, P. K., T. Beer and D. Batten Life cycle assessment of biodiesel production from microalgae in ponds. Bioresource Technology, 102: ) Chand, N Plant oils - Fuel of the future. Journal of Scientific and Industrial Research, 61: ) Chisti, Y Biodiesel from microalgae. Biotechnology Advances, 25: ) Enweremadu, C. C and M. M. Mbarawa Technical aspects of production and analysis of biodiesel from used cooking oil - A review. Renewable and Sustainable Energy Reviews, 13: ) FAO The State of Food and Agriculture, Biofuels: prospects, risks and opportunities. Rome: Food and Agriculture Organization of the United Nations (2008). 12) Gerpen, V Biodiesel processing and production. Fuel Processing Technology, 86: ) Gunvachai, K., M. G. Hassan, G. Shama and C. Hellgardt A new solubility model to describe biodiesel formation kinetics. International Chemical Engineering, 85 (B5): ) Hossain, A. B. M. S., Salleh, A., Boyce, A. N., Chowdhury, P and Naqiuddin, M Biodiesel Fuel Production from Algae as Renewable Energy. American Journal of Biochemistry and Biotechnology, 4 (3):

6 K. Sivakumar/International Journal of Innovations in Agricultural Sciences (IJIAS), 1(1): ) Janaun, J and N. Ellis Perspectives on biodiesel as a sustainable fuel. Renewable and Sustainable Energy Reviews, 14: ) Kansedo, J., K. T. Lee and S. Bhatia Cerbera odollam (sea mango) oil as a promising non-edible feedstock for biodiesel production. Fuel, 88: ) Leung, D. Y. C., X. Wu, M. K. H. Leung A review on biodiesel production using catalyzed transesterification. Applied Energy, 87: ) Mata, T. M., A. A. Martins, N. S. Caetano Microalgae for biodiesel production and other applications: A Review. Renewable and Sustainable Energy Reviews, 14: ) Meher, L. C., D. V. Sagar and S. N. Naik Technical aspects of biodiesel production by transesterification - A review. Renewable and Sustainable Energy Reviews, 10: ) Puppan, D Environmental evaluation of biofuels, Periodica Polytechnica Series. Social and Management Sciences, 10: ) Xu, X., L. Miao, Q. Y. Wu High quality biodiesel production from a microalgae Chlorella protothecoides by heterotrophic growth. Journal of Biotechnology, 126: ) Zabeti, M., W. M. Daud and M. K. Aroua Activity of solid catalysts for biodiesel production: A review. Fuel Processing Technology, 90: How to Cite this Article: K. Sivakumar, G. Kumaresan and K. Kesavamoorthy Production of Biodiesel by Microalgae and Macroalgae from Mangroves of Pichavaram. International Journal of Innovations in Agricultural Sciences, 1 (2): DOI: /ijias Access this Article in Online Quick Response Code Website DOI Number DOI: /ijias

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